人上皮性卵巢癌组织中PFTK1的表达及其对细胞迁移、侵袭及增殖能力的影响

Research of PFTK1 expression in human epithelial ovarian cancer and its effect on migration,invasion and proliferation of epithelial ovarian cancer cells

目的:研究PFTAIRE蛋白激酶1(PFTAIRE protein kinase 1,PFTK1)在人上皮性卵巢癌(epithelial ovarian cancer,EOC)组织中的表达,以及PFTK1基因的沉默对人卵巢癌细胞迁移、侵袭及增殖能力的影响。方法:随机选取我院2015年6月至2016年11月收治的25例EOC患者为研究对象,采用Western blot检测癌组织以及癌旁组织中PFTK1蛋白的表达情况。选取PFTK1高表达的人卵巢癌细胞株SKOV3为研究对象,转染PFTK1-siRNA后,采用流式细胞仪检测PFTK1基因的沉默对SKOV3细胞周期的影响,采用细胞迁移和侵袭实验(Transwell)检测SKOV3细胞迁移、侵袭能力的变化,采用平板克隆形成实验检测SKOV3细胞增殖能力的变化。结果:Western blot检测结果显示,EOC患者癌组织中的PFTK1蛋白表达量显著高于癌旁组织(P<0.01);流式细胞仪检测结果显示,PFTK1-siRNA组细胞出现明显的生长抑制,细胞多停滞在G_(0)/G_(1)期,与对照组相比差异显著(P<0.01);细胞迁移和侵袭实验(Transwell)结果显示,PFTK1-siRNA组细胞的迁移和侵袭能力与对照组比较显著下降,且差异均显著(P<0.01);平板克隆形成实验结果显示,PFTK1-siRNA组细胞增殖能力显著降低,与对照组相比差异显著(P<0.01)。结论:PFTK1在人EOC癌组织中呈现高表达状态;PFTK1基因沉默后,EOC细胞株SKOV3的细胞迁移、侵袭及增殖能力均受到不同程度的抑制,这将为基因治疗人卵巢癌提供一定的理论依据。

Objective:To investigate the expression of PFTAIRE protein kinase 1(PFTK1)in the tissue of human epithelial ovarian cancer(EOC),and the effects of silencing PFTK1 gene on migration,invasion and proliferation of human ovarian cancer cells.Methods:The 25 cases of EOC patients in our hospital from June 2015 to November 2016 were selected as the participants randomly.The expression of PFTK1 protein in cancer tissues and paracancerous tissues was detected by Western blot.And we selected human ovarian cancer cell line SKOV3 with PFTK1 overexpressed as the research object.After transfection with PFTK1-siRNA,the effect of silencing PFTK1 gene on SKOV3 cells cycle was detected by flow cytometry.The abilities of migration and invasion of SKOV3 cells were detected by cell migration and invasion assay(Transwell).At last,the capacity of proliferation of SKOV3 cells was detected by plate clone formation assay.Results:The results of Western blot test showed that the expression of PFTK1 protein in EOC was significantly higher than that in paracancerous tissues(P<0.01).Flow cytometry results showed that cells in PFTK1-siRNA group showed significant growth inhibition,and most of cells stagnated in the G_(0)/G_(1) phase,which was significantly different from that of the control group(P<0.01).The results of cell migration and invasion assay showed that the migration and invasion of cells in PFTK1-siRNA group were significantly decreased compared with control group,and the differences were significant(P<0.01).The results of plate clone formation assay showed that the cell proliferation ability of PFTK1-siRNA group was significantly lower than that of the control group(P<0.01).Conclusion:PFTK1 overexpressed in cancer tissues of human EOC.After PFTK1 gene silencing,the abilities of migration,invasion and proliferation of human EOC cell line SKOV3 are inhibited to different degrees,which will provide a solid theoretical basis for gene therapy for human ovarian cancer.